Rose JB, Crews L, Rockenstein E, Adame A, Mante M, Hersh LB, Gage FH, Spencer B, Potkar R, Marr RA, Masliah E.
Neuropeptide Y fragments derived from neprilysin processing are neuroprotective in a transgenic model of Alzheimer's disease.J Neurosci.
2009 Jan 28;29(4):1115-25.
The comment that the cleavage of neuropeptide Y to generate a biologically active fragment by neprilysin (Neutral EndoPeptidase-24.11) is the first such example for the enzyme is incorrect. At least one example has previously been reported in the metabolism of calcitonin gene-related peptide (CGRP) (Davies et al., 1992).
REAGENTS/MATERIAL: Antibodies used in this study are: Immunocytochemical analysis of neurodegeneration:
monoclonal mouse anti-synaptophysin (Millipore Bioscience);
monoclonal mouse anti-MAP2 (AP20) (Millipore Bioscience);
monoclonal mouse anti-NeuN (A60) (Millipore Bioscience) or
monoclonal mouse anti-GFAP (GA5) (Millipore Bioscience). Western Bloting analysis:
monoclonal mouse anti-FL APP (22C11) (Millipore Bioscience);
monoclonal mouse anti-human APP (8E5) (Elan Pharmaceuticals);
monoclonal mouse anti-Amyloid β (6E10) (Signet Laboratories);
monoclonal mouse anti-NEP (Abcam);
monoclonal mouse anti-BDNF (35928.11) (Calbiochem);
monoclonal mouse anti-NGF (25623.1) (Calbiochem, discontinued);
polyclonal anti-Neurotropin-3 (Promega);
monoclonal mouse anti-Neurotropin-4 (36507) (R&D Systems) and
rabbit anti-Neuropeptide Y (Peninsula Laboratories Bachem) Determination of Aβ, SP, ENK, and NPY levels by ELISA:
ELISA kits for NPY (Phoenix Pharmaceuticals);
Substance P (Assay Designs) and
Met-ENK (Peninsula Laboratories Bachem) Analysis of FL-NPY and NPY CTFs by immunoblot and immunocytochemistry:
rabbit polyclonal anti-FL-NPY (Peninsula Laboratories);
an antibody against NPY CTFs that was prepared at Invitrogen by immunizing rabbits with the peptide corresponding to the amidated 31–36 aa of NPY with an extra Cys added to the N-terminus for conjugation, and
rabbit polyclonal anti-NPY CTFs (20 –36) (Santa Cruz). Double immunocytochemical and laser-scanning confocal analysis:
monoclonal anti-NEP (Abcam);
monoclonal mouse anti-APP (8E5) (Elan Pharmaceuticals) or rabbit polyclonal antibodies against FL-NPY (Peninsula Laboratories). Lentivirus NEP preparation:
Flow cytometry using a monoclonal mouse anti-NEP (56C6) (Research Diagnostics Fitzgerald)